Method of chinese herbal medicine extract used for treating multiple diseases caused by drug resistant bacteria infection

ABSTRACT

Disclosed is a new drug application of a Pithecellobium clypearia Benth Extract (EA), and particularly is a method of the EA used for treating multiple diseases caused by drug resistant bacteria infection. Related drug resistant bacteria include a Multi-Drug Resistant (MDR) Acinetobacter baumannii (MDRAB), an MDR Pseudomonas aeruginosa (MDRPA), an Extended-Spectrum Beta-Lactamase (ESBL) producing Escherichia coli (ECO) and an ESBL-producing Klebsiella pneumonia (KPN).

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is divisional application of U.S. patent application Ser. No. 15/920,480 filed on Mar. 14, 2018, which is a continuation application of PCT application No. PCT/CN2017/071671 filed on Jan. 19, 2017, which claims the benefit of Chinese patent application Nos. 201610308716.7, 201610308717.1, 201610308207.4 and 201610308718.6, each filed on May 10, 2016. The contents of all of the above are hereby incorporated by reference.

TECHNICAL FIELD

The present disclosure relates to a new drug application of a Pithecellobium clypearia Benth Extract (EA), and particularly, to a method of the EA used for treating multiple diseases caused by drug resistant bacteria infection.

BACKGROUND

The twenty-first century is an age of Multi-Drug Resistant (MDR) bacteria. After 60 years of clinical use of an antibiotic, more and more hospital infections and infections with the MDR bacteria have become a major challenge to clinical antibacterial treatment at present. Since a first case of clinical Methicillin-Resistant Staphylococcus Aureus (MRSA) infected patient was reported by Jerons in 1961, the MRSA infections have gradually spread all over the world till now. In 2011, China Bacterial Resistance Surveillance in clinic showed that, in distribution of main drug resistant bacteria, Escherichia coli and Klebsiella pneumonia produced Extended-Spectrum Beta-Lactamase (ESBL) strains respectively were 50.7% and 38.5%, and the transition and current status of the drug resistance thereof are highly concerned. In addition, according to an annual report 2010 from a Drug Resistance Surveillance Cooperating Group of Ministry of Health, pathogenic bacteria separated by Acinetobacter baumannii and Pseudomonas aeruginosa in a hospital ICU ranked the top, and the drug resistance rates of the Acinetobacter baumannii to imipenem (IMP) and meropenem respectively were up to 60.4% and 61.4%. To sum up, in various MRSA and ESBL produced bacteria, the clinical occupied rates of the MDR Acinetobacter baumannii (MDRAB) and Pseudomonas aeruginosa (MDRPA) have been growing every year. Under the pressure that the drug resistance rates are rising continuously, the antibiotic treatment is facing an enormous challenge. In order to prevent further deterioration of a bacterial drug resistant phenomenon, experts and scholars are sparing no effort to discover a new method for inhibiting bacterial growth and treating a bacteria-induced disease. A research has been reported and proved that traditional Chinese herbal medicines such as Coptis chinensis, Scutellaria baicalensis and Forsythia suspensa have a certain inhibition effect to different drug resistant bacteria. The key to further research the Chinese herbal medicine to inhibit the growth of the drug resistant bacteria is to discover a new Chinese herbal medicine with a stronger bactericidal capacity and a wider drug resistant inhibitory spectrum.

With Pithecellobium bigemimum (L.) Benth as a scientific name, the Pithecellobium clypearia Benth is dry young branches and leaves of mimosa pithecellobium plant that is the Pithecellobium clypearia Benth. It is cold, tastes bitter and has the effects of clearing away heat and toxic materials, astringing dampness and healing sore. It is a unique southern medicinal material for treating multiple heat toxin symptoms.

Currently, there has disclosed in a literature that the Pithecellobium clypearia Benth and an extract thereof have an antiviral effect, but lacks a research in an effect of the Pithecellobium clypearia Benth and the extract thereof in aspect of anti-drug resistant bacteria.

SUMMARY

In order to overcome the above defects, the present disclosure discloses an application of an EA in preparing a drug resistant bacteria resistant drug and in an aspect of a sensibilization effect by using with a similar antibiotic thereof, specifically:

1. a method of the EA used for treating disease caused by an MDRAB resistant infection, in which the EA is used with an antibiotic IMP or Tetracycline (TE) or Polymycin B (POLB) or Ceftazidime (CAZ) or Levofloxacin (LVX) and all show an obvious sensibilization effect;

2. a method of the EA used for treating disease caused by an MDRPA resistant infection, in which the EA is used with an antibiotic LVX or IMP or Amikacin (AMK) or CAZ or Cefoperazone (CFP) and all show an obvious sensibilization effect;

3. a method of the EA used for treating disease caused by an ESBL-producing Escherichia coli (ECO) resistant infection, in which the EA is used with an antibiotic AMK or Compound Sulfamethoxazole (SXT) and all show an obvious sensibilization effect;

4. a method of the EA used for treating disease caused by an ESBL-producing Klebsiella pneumonia (KPN) resistant infection, in which the EA shows an obvious inhibitory effect to the ESBL-producing KPN.

The EA preferably is a Pithecellobium clypearia Benth water extract or a Pithecellobium clypearia Benth ethanol extract.

A method for preparing the EA is as follows: extracting Pithecellobium clypearia Benth coarse powder with water or an ethanol aqueous solution having a concentration of 10%-95% by a volume ratio, and then extracting an obtained extracting solution with ethyl acetate, in which the obtained extract is a final product. The ethanol aqueous solution preferably is an ethanol aqueous solution having the concentration of 60% by the volume ratio.

In the present disclosure, the drug includes a human drug or an animal drug, or a soil treatment drug or preparation.

The present disclosure achieves the following beneficial effects.

The present disclosure first discloses the antibacterial effect of the EA to the MDRAB, the MDRPA, the ESBL-producing ECO and the ESBL-producing KPN and the sensibilization effect by using with the antibiotics thereof.

A test proves that the EA by using with the IMP or the TE or the POLB or the CAZ or the LVX has a synergistic effect to the MDRA resistance, and compared with the single use, the usage of the antibiotic is reduced by 50%-87%.

The EA by using with the LVX or the IMP or the AMK or the CAZ or the CFP has a synergistic effect to the MDRPA resistance, and compared with the single use, the usage of the antibiotic is reduced by 50%-99.2%.

The EA by using with the AMK or the SXT has a synergistic effect to the ESBL-producing ECO, and compared with the single use, the usage of the antibiotic is reduced by 75%-99.3%.

The EA in the present disclosure can be taken as a natural antibacterial drug for the above drug resistant bacteria or a sensitizer similar to the antibiotic, and is applied to treatment of a disease caused by the above bacteria. The present disclosure provides a new way and a replacement drug to solve the drug resistant problem of the antibiotics, and is applied to the human drug, other animal drugs and soil remediation of corresponding infectious bacteria. It is a natural plant extract and has no side effect, and the extracting method is simple and environment-friendly.

DETAILED DESCRIPTION OF THE EMBODIMENTS

The EA to resist the drug resistant bacteria and the pharmacological actions of the sensibilization effect to similar antibiotics will be further described with the reference to the following implementation solutions.

Preparation of the EA: the Pithecellobium clypearia Benth is provided by Guangzhou Huacheng Pharmaceutical Factory. An appropriate amount of Pithecellobium clypearia Benth medicinal materials are taken and are crushed into coarse powder; the coarse powder is reflowed for two times with water or with a 10%-95% ethanol aqueous solution, each time for 2 h, and is filtered; filtrates are merged and are condensed to obtain an extractum (namely, the Pithecellobium clypearia Benth water or ethanol extract); after being taken and suspended in the water, the extractum is extracted with ethyl acetate for three times, and ethyl acetate extracting solutions are merged and are condensed to obtain an ethyl acetate extract. The extract obtained by reflowing with a 10% ethanol aqueous solution is referred to as a Pithecellobium clypearia Benth 10% ethanol extract, and the extract obtained by reflowing with other concentrations of ethanol aqueous solutions are by that analogy.

Strains: test strains in the present disclosure all are provided by the Clinical Microbiology Lab of Department of Lab Medicine at First Affiliated Hospital of Sun Yat-Sen University and the drug resistance is tested and confirmed by the same.

Mycoplasma Hominis (MH) broth culture medium: 2.1 g of MH broth dry powder (British OXOID LTD.) is taken and a volume is fixed to 100 ml; the pH is adjusted to 7.0 with NAOH; and the culture medium is sterilized at a high pressure and is placed into a refrigerator at 4° C. for later use.

Determination methods of a Minimal Inhibitory Concentration (MIC) and a Minimum Bactericidal Concentration (MBC) of a test product: the MIC and the MBC of the Pithecellobium clypearia Benth water or ethanol extract to the MDRAB are determined by a microdilution broth method. And it is operated by referring to the microdilution broth method recommended by National Committee for Clinical Laboratory Standards (NCCLS).

Test methods of the sensibilization effects of the test product to similar antibiotics: it is operated by referring to a checkerboard assay recommended by NCCLS.

I. Inhibitory and Bactericidal Tests of MDRA Resistance of the EA and Test of the Sensibilization Effect by Respectively Using with the IMP or the TE or the POLB or the CAZ or the LVX

Embodiment 1

1. Test Method

1) MIC Determination

The EA, the IMP, the TE, the POLB, the CAZ and the LVX are respectively diluted in the MH broth culture medium by a series of ratios, with each pore for 50 μl; the inoculant bacteria are adjusted to 1.0*10⁶ CFU/ml and 50 μl of a bacteria solution is added to each pore; the culture is at 35° C. and for 24 h; and the concentration, at which no precipitate occurs, of a minimum antibacterial drug is the MIC.

2) MBC Determination

A spread plate count method is adopted. 50 μl of bacterial suspension is absorbed from a pore for sterile growth in item 1) to a blood plate, is uniformly coated and is cultured for 24 h at 35° C. Bacterial colonies are counted, and the concentration, at which the initial number of experimental viable bacteria is reduced by 99.9% or more, of the minimum antibacterial drug is the MBC.

By determining the MIC and the MBC of the drug and making statistics of the data to obtain MIC₅₀, MIC₉₀, MBC₅₀ and MBC₉₀, the effect of the MDRA resistance of the drug is evaluated.

3) Checkerboard Assay

The checkerboard assay is performed in a 96-pore sterile culture plate. The EA, the IMP, the TE, the POLB and the CAZ are respectively diluted into a series of concentrations in a doubling manner in the MH broth culture medium, and two drugs are combined respectively at ¼MIC to 4MIC; 25 μl of A drug and 25 μl of B drug are respectively added to each pore, the concentration of the bacterial suspension is adjusted to 1.0*10⁶ CFU/ml, 50 μl of the bacteria solution is inoculated to each pore and is cultured for 24 h at 35° C.; and then, the MIC to the MDRAB after the A drug is used with the B drug is observed.

Calculation of a Fractional Inhibitory Concentration (FIC): FIC=S20b combined-use MIC/S20b single-use MIC+antibiotic combined-use MIC/antibiotic single-use MIC. By calculating the FIC, the combined antibacterial interaction between the EA and the antibiotic is evaluated, with synergistic effect for FIC≤0.5, additive effect for 0.5<FIC≤1, indifferent effect for 1<FIC≤2 and antagonistic effect for FIC>2. According to a sterile growth pore, the optimum concentration ratio between the EA and the antibiotic is founded out. At last, the effect of the EA to strengthen the efficacy of the antibiotic is evaluated.

The test products in the embodiment include: the Pithecellobium clypearia Benth water extract, the Pithecellobium clypearia Benth 10% ethanol extract, the Pithecellobium clypearia Benth 60% ethanol extract and the Pithecellobium clypearia Benth 95% ethanol extract.

The MDRABs are numbered as A1-A20.

2. Test Results

2.1 In-vitro inhibitory test results of the Pithecellobium clypearia Benth water extract and the five antibiotics (the IMP, the TE, the POLB, the CAZ and LVX) to the MDRABs are shown in table A1.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth water extract to the MDRABs are shown in table A2.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract and the five antibiotics to the MDRABs is shown in table A3.

The statistic analysis on MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth water extract to the MDRABs is shown in table A4.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics and distribution statistical results of the FIC values are shown in table A5 and table A6.

The sensibilization effects of the Pithecellobium clypearia Benth water extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables A7-A12.

TABLE A1 In-vitro inhibitory test results of the Pithecellobium clypearia Benth water extract and the five antibiotics to the MDRABs MIC (μg/ml) Strain No. EA IMP TE LVX CAZ POLB A1 600 1 2 0.5 128 8 A2 600 32 256 16 512 2 A3 600 32 256 4 512 2 A4 600 8 8 0.5 256 2 A5 600 32 32 4 64 4 A6 600 32 256 8 >512 8 A7 600 32 512 8 >512 4 A8 600 32 256 4 512 8 A9 600 32 256 4 512 1 A10 600 16 128 32 512 32 A11 600 32 256 4 512 16 A12 600 32 512 32 512 2 A13 600 32 512 32 256 2 A14 600 32 256 16 512 0.5 A15 1200 32 >512 32 256 2 A16 600 32 512 8 512 4 A17 600 16 512 16 256 64 A18 600 32 512 8 512 4 A19 600 32 512 16 256 4 A20 600 32 256 16 512 2 ATCC27853 300 1 8 1 4 1

TABLE A2 In-vitro bactericidal test results of the Pithecellobium clypearia Benth water extract to the MDRABs Strain No. MBC (μg/ml) A1 1200 A2 1200 A3 1200 A4 1200 A5 1200 A6 1200 A7 1200 A8 1200 A9 1200 A10 1200 A11 1200 A12 1200 A13 1200 A14 1200 A15 2400 A16 1200 A17 1200 A18 1200 A19 1200 A20 1200

TABLE A3 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract and the antibiotics to the MDRABs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 600-1200 600 600 IMP 1-32 32 32 TE  2-512 256 512 POLB 0.5-64  4 16 CAZ 64-512 512 512 LVX 0.5-32  8 32

TABLE A4 Bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract to the MDRABs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 1200-2400 1200 1200

TABLE A5 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics Strain FIC No. EA + IMP EA + TE EA + POLB EA + CAZ EA + LVX A1 1 1 1.5 0.75 0.625 A2 1 1 1 1 1.5 A3 1 0.75 1.25 0.75 0.375 A4 0.625 0.75 0.5 0.625 0.375 A5 1.5 0.375 0.375 1 0.5 A6 1 0.5 0.5 1 1 A7 1 1 0.75 1 1 A8 1 1 1.007813 1 1.25 A9 1.5 0.75 1.0625 0.375 0.265625 A10 1 1.5 0.5 1 0.75 A11 1.25 0.75 0.5 1 1 A12 1 0.75 1.03125 0.5 0.5625 A13 1.5 0.5 1.5 1 0.75 A14 0.375 0.375 0.625 0.375 1 A15 0.375 0.3125 0.53125 1 0.501953 A16 0.5 0.625 0.515625 1.125 0.5 A17 0.375 1.25 1.03125 0.625 1 A18 0.375 0.75 1.5 1 0.375 A19 0.5 0.375 1 1 1 A20 0.5 0.5 1 0.375 0.5

TABLE A6 Distribution statistics of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics EA + EA + EA + EA + EA + FIC_Range IMP TE POLB CAZ LVX FIC ≤ 0.5 35% 35% 25% 20% 35% 0.5 < FIC ≤ 1 45% 55% 35% 75% 55% 1 < FIC ≤ 2 20% 10% 40%  5% 10% FIC > 2 — — — — —

TABLE A7 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 1 0.5 A2 600 300 32 16 A3 600 300 32 16 A4 600 300 8 1 A5 600 600 32 16 A6 600 300 32 16 A7 600 300 32 16 A8 600 300 32 16 A9 600 600 32 16 A10 600 300 16 8 A11 600 600 32 8 A12 600 300 32 16 A13 600 600 32 16 A14 600 150 32 4 A15 1200 300 32 4 A16 600 150 32 8 A17 600 150 16 2 A18 600 75 32 8 A19 600 150 32 8 A20 600 150 32 8

TABLE A8 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the TE and MIC after combined use EA EA TE TE Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 2 1 A2 600 300 256 128 A3 600 300 256 64 A4 600 300 8 2 A5 600 150 32 4 A6 600 150 256 64 A7 600 300 512 256 A8 600 300 256 128 A9 600 300 256 64 A10 600 600 128 64 A11 600 300 256 64 A12 600 150 512 256 A13 600 150 512 128 A14 600 150 256 32 A15 1200 300 1024 64 A16 600 300 512 64 A17 600 600 512 128 A18 600 300 512 128 A19 600 150 512 64 A20 600 150 256 64

TABLE A9 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the POLB and MIC after combined use EA EA POLB POLB Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 600 8 4 A2 600 300 2 1 A3 600 600 2 0.5 A4 600 150 2 0.5 A5 600 150 4 0.5 A6 600 150 8 2 A7 600 300 4 1 A8 600 600 8 0.0625 A9 600 600 1 0.0625 A10 600 150 32 8 A11 600 150 16 4 A12 600 600 2 0.0625 A13 600 600 2 1 A14 600 300 0.5 0.0625 A15 1200 600 2 0.0625 A16 600 300 4 0.0625 A17 600 600 64 2 A18 600 600 4 2 A19 600 300 4 2 A20 600 300 2 1

TABLE A10 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 150 128 64 A2 600 300 512 256 A3 600 150 512 256 A4 600 300 256 32 A5 600 300 64 32 A6 600 300 1024 512 A7 600 300 1024 512 A8 600 300 512 256 A9 600 150 512 64 A10 600 300 512 256 A11 600 300 512 256 A12 600 150 512 128 A13 600 300 256 128 A14 600 150 512 64 A15 1200 600 256 128 A16 600 600 512 64 A17 600 300 256 32 A18 600 300 512 256 A19 600 300 256 128 A20 600 150 512 64

TABLE A11 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 0.5 0.0625 A2 600 300 16 16 A3 600 150 16 2 A4 600 150 0.5 0.0625 A5 600 150 4 1 A6 600 300 8 4 A7 600 300 8 4 A8 600 150 4 4 A9 600 150 4 0.0625 A10 600 300 32 8 A11 600 300 4 2 A12 600 300 32 2 A13 600 300 32 8 A14 600 300 16 8 A15 1200 600 32 0.0625 A16 600 150 8 2 A17 600 300 16 8 A18 600 150 8 1 A19 600 300 16 8 A20 600 150 16 4

TABLE A12 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + IMP 8 16 EA + TE 64 128 EA + POLB 1 4 EA + CAZ 128 256 EA + LVX 2 8

With the single use of the Pithecellobium clypearia Benth water extract to the MDRABs, the MIC₅₀ is 600 μg/ml, the MIC₉₀ is 600 μg/ml, the MBC₅₀ is 1200 μg/ml, and the MBC₉₀ is 1200 μg/ml.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth water extract and the IMP that the two drugs have the synergistic effect or partial synergistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the concentration of the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 16 μg/ml and is reduced by 50%.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth water extract and the TE to the 20 MDRABs that the two drugs have the synergistic effect or partial synergistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the TE is reduced from single-use 256 μg/ml to 64 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 128 μg/ml and is reduced by 75%.

For the 20 MDRABs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth water extract and the POLB that the two drugs have no antagonistic effect, wherein 25% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the POLB is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 16 μg/ml to 4 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth water extract and the CAZ to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 512 μg/ml to 128 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth water extract and the LVX to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 35% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRAB, when the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 8 μg/ml to 2 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

2.2. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics (the IMP, the TE, the POLB, the CAZ and the LVX) to the MDRABs are shown in table A13.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRABs are shown in table A14.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics to the MDRABs is shown in table A15.

The statistic analysis on MBC₅₀ and MBC₉₀ of the EA to the MDRABs is shown in table A16.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table A17 and table A18.

The sensibilization effects of the Pithecellobium clypearia Benth 10% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables A19-A24.

TABLE A13 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics to the MDRABs MIC (μg/ml) Strain No. EA IMP TE LVX CAZ POLB A1 600 1 2 0.5 128 8 A2 600 32 256 16 512 2 A3 600 32 256 4 512 2 A4 600 8 8 0.5 256 2 A5 600 32 32 4 64 4 A6 600 32 256 8 >512 8 A7 600 32 512 8 >512 4 A8 600 32 256 4 512 8 A9 600 32 256 4 512 1 A10 600 16 128 32 512 32 A11 600 32 256 4 512 16 A12 600 32 512 32 512 2 A13 600 32 512 32 256 2 A14 600 32 256 16 512 0.5 A15 600 32 >512 32 256 2 A16 600 32 512 8 512 4 A17 600 16 512 16 256 64 A18 600 32 512 8 512 4 A19 600 32 512 16 256 4 A20 600 32 256 16 512 2 ATCC27853 300 1 8 1 4 1

TABLE A14 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRABs Strain No. MBC (μg/ml) A1 1200 A2 1200 A3 1200 A4 1200 A5 1200 A6 1200 A7 1200 A8 1200 A9 1200 A10 1200 A11 1200 A12 1200 A13 1200 A14 1200 A15 1200 A16 1200 A17 1200 A18 1200 A19 1200 A20 1200

TABLE A15 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract and the antibiotics to the MDRABs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 600 600 600 IMP 1-32 32 32 TE  2-512 256 512 POLB 0.5-64  4 16 CAZ 64-512 512 512 LVX 0.5-32  8 32

TABLE A16 bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRABs MBC (μg /ml) Drug Range MBC₅₀ MBC₉₀ EA 1200 1200 1200

TABLE A17 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics Strain FIC No. EA + IMP EA + TE EA + POLB EA + CAZ EA + LVX A1 1 1 1 1 0.625 A2 0.75 0.75 1 1 1 A3 0.75 0.75 1 1 0.625 A4 1.5 0.5 1 0.5 0.625 A5 0.5 0.375 0.75 1 1 A6 0.625 0.75 0.375 1 1 A7 0.5 0.5 0.375 1 1 A8 1 0.5 0.507813 0.75 1 A9 0.375 0.375 0.5625 0.75 0.265625 A10 0.75 0.75 0.375 1 0.75 A11 0.5 0.5 1 0.75 1 A12 0.5 0.5 1 0.75 0.75 A13 0.5 1 1 1 0.75 A14 1 1 0.625 0.375 1 A15 0.75 0.625 1 1 1 A16 0.5 1 0.75 0.5 0.75 A17 0.75 0.375 0.75 0.5 0.5 A18 1 0.5 0.75 1 1 A19 0.5 1 0.75 0.5 0.375 A20 0.5 1.5 1 0.5 0.5

TABLE A18 Distribution statistics of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range IMP TE POLB CAZ LVX FIC ≤ 0.5 45% 45% 15% 30% 20% 0.5 < FIC ≤ 1 50% 50% 85% 70% 80% 1 < FIC ≤ 2  5%  5% — — — FIC > 2 — — — — —

TABLE A19 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 1 0.5 A2 600 300 32 8 A3 600 300 32 8 A4 600 300 8 8 A5 600 150 32 8 A6 600 300 32 4 A7 600 150 32 8 A8 600 300 32 16 A9 600 75 32 8 A10 600 150 16 8 A11 600 150 32 8 A12 600 150 32 8 A13 600 150 32 8 A14 600 300 32 16 A15 600 300 32 8 A16 600 150 32 8 A17 600 300 16 4 A18 600 300 32 16 A19 600 150 32 8 A20 600 150 32 8

TABLE A20 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the TE and MIC after combined use EA EA TE TE Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 2 1 A2 600 300 256 64 A3 600 300 256 64 A4 600 150 8 2 A5 600 150 32 4 A6 600 300 256 64 A7 600 150 512 128 A8 600 150 256 64 A9 600 150 256 32 A10 600 300 128 32 A11 600 150 256 64 A12 600 150 512 128 A13 600 300 512 256 A14 600 300 256 128 A15 600 300 1024 128 A16 600 300 512 256 A17 600 150 512 64 A18 600 150 512 128 A19 600 300 512 256 A20 600 300 256 256

TABLE A21 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the POLB and MIC after combined use EA EA POLB POLB Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 8 4 A2 600 300 2 1 A3 600 300 2 1 A4 600 300 2 1 A5 600 300 4 1 A6 600 150 8 1 A7 600 75 4 1 A8 600 300 8 0.0625 A9 600 300 1 0.0625 A10 600 150 32 4 A11 600 300 16 8 A12 600 300 2 1 A13 600 300 2 1 A14 600 300 0.5 0.0625 A15 600 300 2 1 A16 600 300 4 1 A17 600 300 64 16 A18 600 300 4 1 A19 600 300 4 1 A20 600 300 2 1

TABLE A22 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 128 64 A2 600 300 512 256 A3 600 300 512 256 A4 600 150 256 64 A5 600 300 64 32 A6 600 300 1024 512 A7 600 300 1024 512 A8 600 150 512 256 A9 600 300 512 128 A10 600 300 512 256 A11 600 300 512 128 A12 600 300 512 128 A13 600 300 256 128 A14 600 150 512 64 A15 600 300 256 128 A16 600 150 512 128 A17 600 150 256 64 A18 600 300 512 256 A19 600 150 256 64 A20 600 150 512 128

TABLE A23 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 0.5 0.0625 A2 600 300 16 8 A3 600 300 16 2 A4 600 300 0.5 0.0625 A5 600 300 4 2 A6 600 300 8 4 A7 600 300 8 4 A8 600 300 4 2 A9 600 150 4 0.0625 A10 600 300 32 8 A11 600 300 4 2 A12 600 300 32 8 A13 600 300 32 8 A14 600 300 16 8 A15 600 300 32 16 A16 600 150 8 4 A17 600 150 16 4 A18 600 300 8 4 A19 600 150 16 2 A20 600 150 16 4

TABLE A24 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + IMP 8 16 EA + TE 64 256 EA + POLB 1 4 EA + CAZ 128 256 EA + LVX 4 8

With the single use of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRABs, the MIC₅₀ is 600 μg/ml, the MIC₉₀ is 600 μg/ml, the MBC₅₀ is 1200 μg/ml, and the MBC₉₀ is 1200 μg/ml.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the IMP that the two drugs have the synergistic effect or partial synergistic effect, wherein 45% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 16 μg/ml and is reduced by 50%.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the TE to the 20 MDRABs that the two drugs have the synergistic effect or partial synergistic effect, wherein 45% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the TE is reduced from single-use 256 μg/ml to 64 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

For the 20 MDRABs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the POLB that the two drugs have no antagonistic effect, wherein 15% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the POLB is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 16 μg/ml to 4 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the CAZ to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 30% with FIC≤0.5 have the synergistic effect. When an ethyl acetate extract of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 512 μg/ml to 128 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the LVX to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 25% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 8 μg/ml to 4 μg/ml and is reduced by 50%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

2.3. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics (the IMP, the TE, the POLB, the CAZ and the LVX) to the MDRABs are shown in table A25.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRABs are shown in table A26.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics to the MDRABs is shown in table A27.

The statistic analysis on MBC₅₀ and MBC₉₀ of the EA to the MDRABs is shown in table A28.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table A29 and table A30.

The sensibilization effects of the Pithecellobium clypearia Benth 60% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables A31-A36.

TABLE A25 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics to the MDRABs MIC (μg/ml) Strain No. EA IMP TE LVX CAZ POLB A1 300 1 2 0.5 128 8 A2 300 32 256 16 512 2 A3 600 32 256 4 512 2 A4 600 8 8 0.5 256 2 A5 300 32 32 4 64 4 A6 300 32 256 8 >512 8 A7 300 32 512 8 >512 4 A8 300 32 256 4 512 8 A9 600 32 256 4 512 1 A10 300 16 128 32 512 32 A11 600 32 256 4 512 16 A12 300 32 512 32 512 2 A13 300 32 512 32 256 2 A14 600 32 256 16 512 0.5 A15 600 32 >512 32 256 2 A16 300 32 512 8 512 4 A17 300 16 512 16 256 64 A18 300 32 512 8 512 4 A19 300 32 512 16 256 4 A20 300 32 256 16 512 2 ATCC27853 300 1 8 1 4 1

TABLE A26 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRABs Strain No. MBC (μg/ml) A1 600 A2 600 A3 1200 A4 1200 A5 600 A6 600 A7 600 A8 1200 A9 600 A10 600 A11 600 A12 600 A13 600 A14 1200 A15 1200 A16 1200 A17 600 A18 600 A19 600 A20 600

TABLE A27 Statistics on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract and the antibiotics to the MDRABs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 300-600  300 600 IMP 1-32 32 32 TE  2-512 256 512 POLB 0.5-64  4 16 CAZ 64-512 512 512 LVX 0.5-32  8 32

TABLE A28 Bactericidal MBC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRABs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 600-1200 600 1200

TABLE A29 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics Strain FIC No. EA + IMP EA + TE EA + POLB EA + CAZ EA + LVX A1 0.75 0.75 1.5 0.75 0.375 A2 0.5 0.5 1.25 0.75 1.25 A3 0.375 0.375 1 0.5 1.0040 A4 0.1875 0.625 0.75 0.5 0.625 A5 0.5 0.75 1 0.75 1.25 A6 0.625 0.75 0.5 1 1 A7 0.625 0.75 0.75 0.75 1 A8 0.625 0.625 1.0080 0.75 1.25 A9 0.375 0.375 0.5625 0.625 0.5156 A10 0.75 0.75 0.5 0.75 1.0020 A11 0.375 0.375 0.5 0.625 1.125 A12 0.3125 0.5 1.25 0.75 1.03125 A13 0.5 0.5 1.5 1 1.0020 A14 0.25 0.375 0.625 0.625 0.625 A15 0.375 0.3125 0.53125 0.75 0.5020 A16 0.5 0.5 1.0156 0.75 0.75 A17 0.625 1 1.03125 0.75 0.75 A18 0.5 0.625 1.125 0.75 0.75 A19 0.375 0.5 1.125 1 1 A20 0.5 0.625 1.25 1 1

TABLE A30 Distribution statistics of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range IMP TE POLB CAZ LVX FIC ≤ 0.5 70% 50% 15% 10%  5% 0.5 < FIC ≤ 1 30% 50% 35% 85% 55% 1 < FIC ≤ 2 — — 50%  5% 40% FIC > 2 — — — — —

TABLE A31 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 300 75 1 0.5 A2 300 75 32 8 A3 600 75 32 8 A4 600 75 8 0.5 A5 300 75 32 8 A6 300 37.5 32 16 A7 300 37.5 32 16 A8 300 37.5 32 16 A9 600 150 32 4 A10 300 75 16 8 A11 600 150 32 4 A12 300 18.75 32 8 A13 300 75 32 8 A14 600 75 32 4 A15 600 150 32 4 A16 300 75 32 8 A17 300 150 16 2 A18 300 75 32 8 A19 300 37.5 32 8 A20 300 75 32 8

TABLE A32 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the TE and MIC after combined use EA EA TE TE Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 300 75 2 1 A2 300 75 256 64 A3 600 150 256 32 A4 600 300 8 1 A5 300 150 32 8 A6 300 75 256 128 A7 300 150 512 128 A8 300 150 256 32 A9 600 150 256 32 A10 300 150 128 32 A11 600 150 256 32 A12 300 75 512 128 A13 300 75 512 128 A14 600 150 256 32 A15 600 150 1024 64 A16 300 75 512 128 A17 300 150 512 256 A18 300 150 512 64 A19 300 75 512 128 A20 300 150 256 32

TABLE A33 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the POLB and MIC after combined use EA EA POLB POLB Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 300 300 8 4 A2 300 300 2 0.5 A3 600 300 2 1 A4 600 150 2 1 A5 300 150 4 2 A6 300 75 8 2 A7 300 75 4 2 A8 300 300 8 0.0625 A9 600 300 1 0.0625 A10 300 75 32 8 A11 600 150 16 4 A12 300 300 2 0.5 A13 300 300 2 1 A14 600 300 0.5 0.0625 A15 600 300 2 0.0625 A16 300 300 4 0.0625 A17 300 300 64 2 A18 300 300 4 0.5 A19 300 300 4 0.5 A20 300 300 2 0.5

TABLE A34 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 300 75 128 64 A2 300 150 512 128 A3 600 150 512 128 A4 600 150 256 64 A5 300 75 64 32 A6 300 150 1024 512 A7 300 150 1024 256 A8 300 75 512 256 A9 600 300 512 64 A10 300 150 512 128 A11 600 300 512 64 A12 300 150 512 128 A13 300 150 256 128 A14 600 75 512 256 A15 600 150 256 128 A16 300 150 512 128 A17 300 75 256 128 A18 300 150 512 128 A19 300 150 256 128 A20 300 150 512 256

TABLE A35 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 300 75 0.5 0.0625 A2 300 75 16 16 A3 300 300 16 0.0625 A4 600 300 0.5 0.0625 A5 300 75 4 4 A6 300 150 8 4 A7 300 150 8 4 A8 300 75 4 4 A9 600 300 4 0.0625 A10 300 300 32 0.0625 A11 600 75 4 4 A12 300 300 32 1 A13 300 300 32 0.0625 A14 600 75 16 8 A15 600 300 32 0.0625 A16 300 75 8 4 A17 300 150 16 4 A18 300 75 8 4 A19 300 150 16 8 A20 300 150 16 8

TABLE A36 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + IMP 8 16 EA + TE 32 128 EA + POLB 0.5 4 EA + CAZ 128 256 EA + LVX 4 8

With the single use of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRABs, the MIC₅₀ is 300 μg/ml, the MIC₉₀ is 600 μg/ml, the MBC₅₀ is 600 μg/ml, and the MBC₉₀ is 1200 μg/ml.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the IMP that the two drugs have the synergistic effect or partial synergistic effect, wherein 70% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 16 μg/ml and is reduced by 50%.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the TE to the 20 MDRABs that the two drugs have the synergistic effect or partial synergistic effect, wherein 50% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the TE is reduced from single-use 256 μg/ml to 32 μg/ml and is reduced by 87.5%; and the MIC₉₀ is reduced from 512 μg/ml to 128 μg/ml and is reduced by 75%.

For the 20 MDRABs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the POLB that the two drugs have no antagonistic effect, wherein 15% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the POLB is reduced from single-use 4 μg/ml to 0.5 μg/ml and is reduced by 87.5%; and the MIC₉₀ is reduced from 16 μg/ml to 4 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the CAZ to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 10% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to ½ of the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 512 μg/ml to 128 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the LVX to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 5% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 8 μg/ml to 4 μg/ml and is reduced by 50%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

2.4. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics (the IMP, the TE, the POLB, the CAZ and the LVX) to the MDRABs are shown in table A37.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRABs are shown in table A38.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics to the MDRABs is shown in table A39.

The statistic analysis on MBC₅₀ and MBC₉₀ of the EA to the MDRABs is shown in table A40.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table A41 and table A42.

The sensibilization effects of the Pithecellobium clypearia Benth 95% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables A43-A48.

TABLE A37 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics to the MDRABs MIC (μg/ml) Strain No. EA IMP TE LVX CAZ POLB A1 600 1 2 0.5 128 8 A2 600 32 256 16 512 2 A3 600 32 256 4 512 2 A4 600 8 8 0.5 256 2 A5 600 32 32 4 64 4 A6 600 32 256 8 >512 8 A7 600 32 512 8 >512 4 A8 600 32 256 4 512 8 A9 600 32 256 4 512 1 A10 600 16 128 32 512 32 A11 600 32 256 4 512 16 A12 600 32 512 32 512 2 A13 600 32 512 32 256 2 A14 600 32 256 16 512 0.5 A15 1200 32 >512 32 256 2 A16 600 32 512 8 512 4 A17 600 16 512 16 256 64 A18 600 32 512 8 512 4 A19 600 32 512 16 256 4 A20 600 32 256 16 512 2 ATCC27853 300 1 8 1 4 1

TABLE A38 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRABs Strain No. MBC (μg/ml) A1 1200 A2 1200 A3 1200 A4 1200 A5 1200 A6 1200 A7 1200 A8 1200 A9 1200 A10 1200 A11 1200 A12 1200 A13 1200 A14 1200 A15 1200 A16 1200 A17 1200 A18 1200 A19 1200 A20 1200

TABLE A39 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract and the antibiotics to the MDRABs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 600-1200 600 600 IMP 1-32 32 32 TE  2-512 256 512 POLB 0.5-64  4 16 CAZ 64-512 512 512 LVX 0.5-32  8 32

TABLE A40 Bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRABs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 1200 1200 1200

TABLE A41 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics Strain FIC No. EA + IMP EA + TE EA + POLB EA + CAZ EA + LVX A1 0.75 0.75 1 1 1 A2 0.5 0.5 0.75 0.75 1 A3 0.5 0.375 1 0.75 0.625 A4 0.3125 0.625 1 0.75 0.375 A5 0.75 0.5 1 1 1 A6 0.75 1 0.5 1 1 A7 0.75 0.75 0.75 0.5 1 A8 0.75 0.625 0.625 1 1 A9 0.375 0.625 0.75 1 0.515625 A10 0.75 0.75 0.5 1 0.3125 A11 0.375 0.625 0.5 0.625 1.5 A12 0.5 0.75 1 0.75 0.53125 A13 0.75 0.5 1 0.75 0.5625 A14 0.75 0.375 1 1 1 A15 0.75 0.3125 0.28125 0.75 0.3125 A16 0.5 0.75 0.515625 1 0.75 A17 0.375 0.75 0.5625 1 0.5 A18 0.5 0.75 0.75 0.5 1 A19 0.5 0.75 0.75 0.75 1 A20 0.5 0.625 0.75 1 0.75

TABLE A42 Distribution statistics of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range IMP TE POLB CAZ LVX FIC ≤ 0.5 55% 30% 20% 10% 20% 0.5 < FIC ≤ 1 45% 70% 80% 90% 75% 1 < FIC ≤ 2 — — — —  5% FIC > 2 — — — — —

TABLE A43 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 150 1 0.5 A2 600 150 32 8 A3 600 150 32 8 A4 600 150 8 0.5 A5 600 300 32 8 A6 600 150 32 16 A7 600 150 32 16 A8 600 150 32 16 A9 600 150 32 4 A10 600 150 16 8 A11 600 150 32 4 A12 600 150 32 8 A13 600 300 32 8 A14 600 300 32 8 A15 1200 600 32 8 A16 600 150 32 8 A17 600 150 16 2 A18 600 150 32 8 A19 600 150 32 8 A20 600 150 32 8

TABLE A44 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the TE and MIC after combined use EA EA TE TE Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 150 2 1 A2 600 150 256 64 A3 600 150 256 32 A4 600 300 8 1 A5 600 150 32 8 A6 600 300 256 128 A7 600 300 512 128 A8 600 300 256 32 A9 600 300 256 32 A10 600 300 128 32 A11 600 300 256 32 A12 600 300 512 128 A13 600 150 512 128 A14 600 150 256 32 A15 1200 300 1024 64 A16 600 300 512 128 A17 600 300 512 128 A18 600 300 512 128 A19 600 300 512 128 A20 600 300 256 32

TABLE A45 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the POLB and MIC after combined use EA EA POLB POLB Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 8 4 A2 600 300 2 0.5 A3 600 300 2 1 A4 600 300 2 1 A5 600 300 4 2 A6 600 150 8 2 A7 600 150 4 2 A8 600 300 8 1 A9 600 300 1 0.25 A10 600 150 32 8 A11 600 150 16 4 A12 600 300 2 1 A13 600 300 2 1 A14 600 300 0.5 0.25 A15 1200 300 2 0.0625 A16 600 300 4 0.0625 A17 600 300 64 4 A18 600 300 4 1 A19 600 300 4 1 A20 600 300 2 0.5

TABLE A46 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 128 64 A2 600 300 512 128 A3 600 300 512 128 A4 600 300 256 64 A5 600 300 64 32 A6 600 300 1024 512 A7 600 150 1024 256 A8 600 300 512 256 A9 600 300 512 256 A10 600 300 512 256 A11 600 300 512 64 A12 600 300 512 128 A13 600 150 256 128 A14 600 300 512 256 A15 1200 300 256 128 A16 600 300 512 256 A17 600 300 256 128 A18 600 150 512 128 A19 600 150 256 128 A20 600 300 512 256

TABLE A47 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) A1 600 300 0.5 0.25 A2 600 300 16 8 A3 600 300 16 2 A4 600 150 0.5 0.0625 A5 600 300 4 2 A6 600 300 8 4 A7 600 300 8 4 A8 600 300 4 2 A9 600 300 4 0.0625 A10 600 150 32 2 A11 600 300 4 4 A12 600 300 32 1 A13 600 300 32 2 A14 600 300 16 8 A15 1200 300 32 2 A16 600 150 8 4 A17 600 150 16 4 A18 600 300 8 4 A19 600 300 16 8 A20 600 150 16 8

TABLE A48 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + IMP 8 16 EA + TE 32 128 EA + POLB 1 4 EA + CAZ 128 256 EA + LVX 4 8

With the single use of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRABs, the MIC₅₀ is 600 μg/ml, the MIC₉₀ is 600 μg/ml, the MBC₅₀ is 1200 μg/ml, and the MBC₉₀ is 1200 μg/ml.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the IMP that the two drugs have the synergistic effect or partial synergistic effect, wherein 55% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 16 μg/ml and is reduced by 50%.

It is indicated by FIC≤1 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the TE to the 20 MDRABs that the two drugs have the synergistic effect or partial synergistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the TE is reduced from single-use 256 μg/ml to 32 μg/ml and is reduced by 87.5%; and the MIC₉₀ is reduced from 512 μg/ml to 128 μg/ml and is reduced by 75%.

For the 20 MDRABs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the POLB that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the POLB is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 16 μg/ml to 4 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the CAZ to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 10% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 512 μg/ml to 128 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the LVX to the 20 MDRABs that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRABs, when the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 8 μg/ml to 4 μg/ml and is reduced by 50%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

II. Inhibitory and Bactericidal Tests of the MDRPA Resistance of the EA and Test of the Sensibilization Effect by Respectively Using with the LVX, the IMP, the AMK, the CAZ and the CFP

Embodiment 2

1. Test Method: The MDRPA (Serial No.: P1-P20) Strains are Tested and are Evaluated with Reference to the Method in the First Embodiment.

2. Test Results:

2.1. In-vitro inhibitory test results of the Pithecellobium clypearia Benth water extract and the five antibiotics (the LVX, the IMP, the AMK, the CAZ and the CFP) to the MDRPAs are shown in table P1.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth water extract to the MDRPAs are shown in table P2.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the EA and the five antibiotics to the MDRPAs is shown in table P3.

The statistic analysis on MBC₅₀ and MBC₉₀ of the EA to the MDRPAs is shown in table P4.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics and distribution statistical results of the FIC values are shown in table P5 and table P6.

The sensibilization effects of the Pithecellobium clypearia Benth water extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables P7-P12.

TABLE P1 In-vitro inhibitory test results of the Pithecellobium clypearia Benth water extract and the five antibiotics to the MDRPAs MIC(μg/ml) Strain No. EA LVX IMP AMK CAZ CFP P1 800 32 32 1 32 128 P2 800 4 64 1 64 256 P3 800 8 64 4 4 16 P4 800 8 32 1 256 8 P5 800 1 64 2 64 512 P6 1600 0.5 32 256 512 512 P7 800 32 32 1 64 512 P8 1600 1 64 1 256 16 P9 800 4 32 1 8 64 P10 800 128 64 4 4 8 P11 800 2 8 1 2 8 P12 800 1 32 32 64 512 P13 800 2 8 2 16 256 P14 800 32 32 2 16 8 P15 800 2 32 2 256 64 P16 1600 32 16 256 32 >512 P17 800 4 16 1 32 16 P18 800 4 32 16 64 256 P19 800 16 64 1 4 64 P20 800 4 4 2 256 >512 ATCC27853 400 1 4 2 4 8

TABLE P2 In-vitro bactericidal test results of the Pithecellobium clypearia Benth water extract to the MDRPAs Strain No. MBC (μg/ml) P1 1600 P2 1600 P3 1600 P4 1600 P5 1600 P6 1600 P7 1600 P8 1600 P9 1600 P10 1600 P11 1600 P12 1600 P13 1600 P14 3200 P15 1600 P16 1600 P17 1600 P18 1600 P19 3200 P20 1600

TABLE P3 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract and the five antibiotics to the MDRPAs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 800-1600  800 1600 CAZ 2-512 32 256 CFP 8-512 64 512 AMK 1-256 1 32 IMP 4-64  32 64 LVX 0.5-128  4 32

TABLE P4 Bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth water extract to the MDRPAs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 1600-3200 1600 1600

TABLE P5 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics FIC Strain EA + EA + EA + EA + EA + No. CAZ CFP AMK IMP LVX P1 0.5 0.375 0.5625 1 0.625 P2 0.5 1 0.1875 0.28125 0.507813 P3 0.75 0.375 0.375 0.28125 0.503906 P4 0.5 1 1.5 1.0625 0.75 P5 1 0.75 1.03125 0.5 0.53125 P6 0.625 0.75 0.75 0.5625 0.3125 P7 0.75 0.53125 0.5625 1.0625 1.000977 P8 0.375 0.15625 0.75 0.28125 0.28125 P9 1 1 0.5625 1 1 P10 1 0.5625 0.375 0.5 0.375 P11 1 0.75 1 0.75 1 P12 0.53125 0.250977 0.3125 0.5625 1 P13 0.5 0.375 0.28125 0.5 1 P14 1 0.5625 0.53125 0.625 1.03125 P15 1 0.5 0.53125 0.5 1.5 P16 0.5 0.375 0.250244 0.375 0.5625 P17 0.5 0.53125 0.5625 0.75 2 P18 0.5 0.75 0.5 0.625 2.03125 P19 1 0.507813 0.5625 0.5625 0.75 P20 0.5 0.500488 0.625 1 0.75

TABLE P6 Distribution statistical results of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range CAZ CFP AMK IMP LVX FIC ≤ 0.5 45% 35% 35% 40% 15% 0.5 < FIC ≤ 1 55% 65% 55% 50% 60% 1 < FIC ≤ 2 — — 10% 10% 25% FIC > 2 — — — — —

TABLE P7 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 200 32 8 P2 800 200 64 16 P3 800 200 4 2 P4 800 200 256 64 P5 800 400 64 32 P6 1600 200 512 256 P7 800 200 64 32 P8 1600 200 256 64 P9 800 400 8 4 P10 800 400 4 2 P11 800 400 2 1 P12 800 400 64 2 P13 800 200 16 4 P14 800 400 16 8 P15 800 400 256 128 P16 1600 400 32 8 P17 800 200 32 8 P18 800 200 64 16 P19 800 400 4 2 P20 800 200 256 64

TABLE P8 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the CFP and MIC after combined use EA EA CFP CFP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 100 128 32 P2 800 400 256 128 P3 800 200 16 2 P4 800 400 8 4 P5 800 200 512 256 P6 1600 400 512 256 P7 800 400 512 16 P8 1600 200 16 0.5 P9 800 400 64 32 P10 800 400 8 0.5 P11 800 200 8 4 P12 800 200 512 0.5 P13 800 200 256 32 P14 800 400 8 0.5 P15 800 200 64 16 P16 1600 200 1024 256 P17 800 400 16 0.5 P18 800 200 256 128 P19 800 400 64 0.5 P20 800 400 1024 0.5

TABLE P9 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the AMK and MIC after combined use EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 1 0.0625 P2 800 100 1 0.0625 P3 800 100 4 1 P4 800 800 1 0.5 P5 800 800 2 0.0625 P6 1600 800 256 64 P7 800 400 1 0.0625 P8 1600 400 1 0.5 P9 800 400 1 0.0625 P10 800 100 4 1 P11 800 400 1 0.5 P12 800 50 32 8 P13 800 200 2 0.0625 P14 800 400 2 0.0625 P15 800 400 2 0.0625 P16 1600 400 256 0.0625 P17 800 400 1 0.0625 P18 800 200 16 4 P19 800 400 1 0.0625 P20 800 100 2 1

TABLE P10 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 16 P2 800 200 64 2 P3 800 200 64 2 P4 800 800 32 2 P5 800 200 64 16 P6 1600 800 32 2 P7 800 800 32 2 P8 1600 400 64 2 P9 800 400 32 16 P10 800 200 64 16 P11 800 200 8 4 P12 800 400 32 2 P13 800 200 8 2 P14 800 400 32 4 P15 800 200 32 8 P16 1600 400 16 2 P17 800 400 16 4 P18 800 400 32 4 P19 800 400 64 4 P20 800 400 4 2

TABLE P11 Sensibilization effect of the Pithecellobium clypearia Benth water extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 4 P2 800 400 4 0.03125 P3 800 400 8 0.03125 P4 800 200 8 4 P5 800 400 1 0.03125 P6 1600 400 0.5 0.03125 P7 800 800 32 0.03125 P8 1600 400 1 0.03125 P9 800 400 4 2 P10 800 100 128 32 P11 800 400 2 1 P12 800 400 1 0.5 P13 800 400 2 1 P14 800 800 32 1 P15 800 800 2 1 P16 1600 800 32 2 P17 800 800 4 4 P18 800 25 4 8 P19 800 400 16 4 P20 800 400 4 1

TABLE P12 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + CAZ 8 64 EA + CFP 0.5 256 EA + AMK 0.0625 2 EA + IMP 2 16 EA + LVX 1 8

With the single use of the EA to the MDRPA, the MIC₅₀ is 800 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 1600 μg/ml, and the MBC₉₀ is 1600 μg/ml.

For the 20 MDRPAs, it is indicated by FIC≤2 with the combined use of the EA and the CAZ that the two drugs have no antagonistic effect, wherein 45% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the EA is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 256 μg/ml to 64 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the EA and the CFP to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the EA is smaller than or equal to the single-use MIC, the MIC₅₀ of the CFP is reduced from single-use 64 μg/ml to 0.5 μg/ml and is reduced by 99.2%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the EA and the AMK that the two drugs have no antagonistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the EA is smaller than or equal to the single-use MIC, the MIC₅₀ of the AMK is reduced from single-use 1 μg/ml to 0.0625 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 32 μg/ml to 2 μg/ml and is reduced by 93.75%.

It is indicated by FIC≤2 with the combined use of the EA and the IMP that the two drugs have no antagonistic effect, wherein 40% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the EA is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 2 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the EA and the LVX to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 15% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRPA, when the concentration of the EA is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

2.2. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics (the LVX, the IMP, the AMK, the CAZ and the CFP) to the MDRPAs are shown in table P13.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRPAs are shown in table P14.

According to statistic analysis, the in-vitro inhibitory and bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics to the MDRPAs are shown in table P15.

According to statistic analysis, the bactericidal MBC₅₀ and MBC₉₀ of the EA to the MDRPAs are shown in table P16.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table P17 and table P18.

The sensibilization effects of the Pithecellobium clypearia Benth 10% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables P19-P24.

TABLE P13 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics to the MDRPAs MIC(μg/ml) Strain No. EA LVX IMP AMK CAZ CFP P1 800 32 32 1 32 128 P2 800 4 64 1 64 256 P3 800 8 64 4 4 16 P4 800 8 32 1 256 8 P5 800 1 64 2 64 512 P6 800 0.5 32 256 512 512 P7 800 32 32 1 64 512 P8 1600 1 64 1 256 16 P9 800 4 32 1 8 64 P10 800 128 64 4 4 8 P11 800 2 8 1 2 8 P12 800 1 32 32 64 512 P13 800 2 8 2 16 256 P14 800 32 32 2 16 8 P15 800 2 32 2 256 64 P16 1600 32 16 256 32 >512 P17 800 4 16 1 32 16 P18 800 4 32 16 64 256 P19 800 16 64 1 4 64 P20 800 4 4 2 256 >512 ATCC27853 400 1 4 2 4 8

TABLE P14 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRPAs Strain No. MBC (μg/ml) P1 1600 P2 1600 P3 1600 P4 1600 P5 1600 P6 1600 P7 1600 P8 1600 P9 1600 P10 1600 P11 1600 P12 1600 P13 1600 P14 1600 P15 1600 P16 1600 P17 1600 P18 1600 P19 3200 P20 1600

TABLE P15 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract and the antibiotics to the MDRPAs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 800-1600  800 800 CAZ 2-512 32 256 CFP 8-512 64 512 AMK 1-256 1 32 IMP 4-64  32 64 LVX 0.5-128  4 32

TABLE P16 Bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRPAs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 1600-3200 1600 1600

TABLE P17 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics Strain FIC No. EA + CAZ EA + CFP EA + AMK EA + IMP EA + LVX P1 0.75 0.5 0.5625 0.75 0.625 P2 0.5 1 0.3125 0.28125 0.507813 P3 0.75 0.375 0.75 0.28125 0.503906 P4 0.5 1 1.5 1 1 P5 0.75 1 1.03125 0.5 0.53125 P6 0.75 0.625 0.75 0.5625 0.3125 P7 0.75 0.515625 0.5625 0.5625 1.000977 P8 0.375 0.15625 0.75 0.28125 0.28125 P9 1 1 0.5625 1 1 P10 0.75 0.75 0.5 0.5 0.75 P11 1 0.75 0.75 0.75 1 P12 0.53125 0.253906 0.5 0.5625 1 P13 0.5 0.375 0.28125 0.5 1 P14 1 0.53125 0.53125 0.625 0.5625 P15 1 0.375 0.53125 0.375 1 P16 0.5 0.375 0.257813 0.375 0.375 P17 0.5 0.53125 0.5625 0.625 1 P18 1 1 0.5 0.375 1.5 P19 1 0.507813 0.5625 0.375 0.75 P20 0.5 0.500977 50.5 1 0.75

TABLE 18 Distribution statistical results of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range CAZ CFP AMK IMP LVX FIC ≤ 0.5 35% 35% 30% 50% 15% 0.5 < FIC ≤ 1 65% 65% 55% 50% 75% 1 < FIC ≤ 2 — — 15% — 10% FIC > 2 — — — — —

TABLE P19 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 200 32 16 P2 800 200 64 16 P3 800 200 4 2 P4 800 200 256 64 P5 800 200 64 32 P6 800 200 512 256 P7 800 200 64 32 P8 1600 200 256 64 P9 800 400 8 4 P10 800 200 4 2 P11 800 400 2 1 P12 800 400 64 2 P13 800 200 16 4 P14 800 400 16 8 P15 800 400 256 128 P16 1600 400 32 8 P17 800 200 32 8 P18 800 400 64 32 P19 800 400 4 2 P20 800 200 256 64

TABLE P20 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the CFP and MIC after combined use EA EA CFP CFP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 200 128 32 P2 800 400 256 128 P3 800 200 16 2 P4 800 400 8 4 P5 800 400 512 256 P6 1600 200 512 256 P7 800 400 512 8 P8 1600 200 16 0.5 P9 800 400 64 32 P10 800 400 8 2 P11 800 200 8 4 P12 800 200 512 2 P13 800 200 256 32 P14 800 400 8 0.25 P15 800 200 64 8 P16 1600 200 1024 256 P17 800 400 16 0.5 P18 800 400 256 128 P19 800 400 64 0.5 P20 800 400 1024 1

TABLE P21 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the AMK and MIC after combined use EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 1 0.0625 P2 800 200 1 0.0625 P3 800 200 4 2 P4 800 800 1 0.5 P5 800 800 2 0.0625 P6 1600 800 256 64 P7 800 400 1 0.0625 P8 1600 400 1 0.5 P9 800 400 1 0.0625 P10 800 200 4 1 P11 800 400 1 0.25 P12 800 200 32 8 P13 800 200 2 0.0625 P14 800 400 2 0.0625 P15 800 400 2 0.0625 P16 1600 400 256 2 P17 800 400 1 0.0625 P18 800 200 16 4 P19 800 400 1 0.0625 P20 800 40000 2 1

TABLE P22 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 200 32 16 P2 800 200 64 2 P3 800 200 64 2 P4 800 400 32 16 P5 800 200 64 16 P6 1600 800 32 2 P7 800 400 32 2 P8 1600 400 64 2 P9 800 400 32 16 P10 800 200 64 16 P11 800 200 8 4 P12 800 400 32 2 P13 800 200 8 2 P14 800 400 32 4 P15 800 200 32 4 P16 1600 400 16 2 P17 800 400 16 2 P18 800 200 32 4 P19 800 200 64 8 P20 800 400 4 2

TABLE P23 Sensibilization effect of the Pithecellobium clypearia Benth 10% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 4 P2 800 400 4 0.03125 P3 800 400 8 0.03125 P4 800 400 8 4 P5 800 400 1 0.03125 P6 1600 400 0.5 0.03125 P7 800 800 32 0.03125 P8 1600 400 1 0.03125 P9 800 400 4 2 P10 800 400 128 32 P11 800 400 2 1 P12 800 400 1 0.5 P13 800 400 2 1 P14 800 400 32 2 P15 800 400 2 1 P16 1600 400 32 4 P17 800 400 4 2 P18 800 400 4 4 P19 800 400 16 4 P20 800 400 4 1

TABLE P24 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + CAZ 16 64 EA + CFP 4 256 EA + AMK 0.0625 4 EA + IMP 2 16 EA + LVX 1 4

With the single use of the Pithecellobium clypearia Benth 10% ethanol extract to the MDRPA, the MIC₅₀ is 800 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 1600 μg/ml, and the MBC₉₀ is 1600 μg/ml.

For the 20 MDRPAs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the CAZ that the two drugs have no antagonistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 32 μg/ml to 16 μg/ml and is reduced by 50%; and the MIC₉₀ is reduced from 256 μg/ml to 64 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the CFP to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 35% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CFP is reduced from single-use 64 μg/ml to 4 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 30% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the AMK is reduced from single-use 1 μg/ml to 0.0625 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 32 μg/ml to 4 μg/ml and is reduced by 87.5%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the IMP that the two drugs have no antagonistic effect, wherein 50% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 2 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the LVX to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 15% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRPA, when the concentration of the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 4 μg/ml and is reduced by 87.5%.

2.3. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics (the LVX, the IMP, the AMK, the CAZ and the CFP) to the MDRPAs are shown in table P25.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRPAs are shown in table P26.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics to the MDRPAs is shown in table P27.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the EA to the MDRPAs is shown in table P28.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table P29 and table P30.

The sensibilization effects of the Pithecellobium clypearia Benth 60% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables P31-P36.

TABLE P25 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics to the MDRPAs MIC(μg/ml) Strain No. EA LVX IMP AMK CAZ CFP P1 400 32 32 1 32 128 P2 200 4 64 1 64 256 P3 400 8 64 4 4 16 P4 400 8 32 1 256 8 P5 400 1 64 2 64 512 P6 400 0.5 32 256 512 512 P7 400 32 32 1 64 512 P8 800 1 64 1 256 16 P9 400 4 32 1 8 64 P10 400 128 64 4 4 8 P11 400 2 8 1 2 8 P12 400 1 32 32 64 512 P13 400 2 8 2 16 256 P14 800 32 32 2 16 8 P15 400 2 32 2 256 64 P16 800 32 16 256 32 >512 P17 400 4 16 1 32 16 P18 400 4 32 16 64 256 P19 800 16 64 1 4 64 P20 800 4 4 2 256 >512 ATCC27853 400 1 4 2 4 8

TABLE P26 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRPAs Strain No. MBC (μg/ml) P1 400 P2 400 P3 800 P4 800 P5 800 P6 800 P7 800 P8 800 P9 1600 P10 1600 P11 1600 P12 1600 P13 1600 P14 1600 P15 1600 P16 1600 P17 1600 P18 1600 P19 1600 P20 1600

TABLE P27 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics to the MDRPAs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 200-800  400 800 CAZ 2-512 32 256 CFP 8-512 64 512 AMK 1-256 1 32 IMP 4-64  32 64 LVX 0.5-128  4 32

TABLE P28 Bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRPAs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 400-1600 1600 1600

TABLE P29 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics Strain FIC No. EA + CAZ EA + CFP EA + AMK EA + IMP EA + LVX P1 0.375 0.375 1.0625 0.625 0.25 P2 0.625 0.75 0.5625 0.53125 0.5078 P3 1.125 1.03125 0.5 0.15625 1.0039 P4 0.375 1.0625 1.5 1.0625 1 P5 1.0078 0.625 1.03125 0.75 1.03125 P6 0.75 1 1.125 1.0625 1.0625 P7 1 0.625 1.0625 1.0625 1.0010 P8 0.3125 0.53125 0.5625 0.53125 0.53125 P9 1.0625 0.625 0.5625 0.75 1 P10 1.125 1.0625 0.375 0.75 0.5 P11 1.25 1.0625 0.75 1.25 2.0625 P12 0.5078 0.5010 0.375 0.5625 1.5 P13 0.75 0.375 0.53125 0.5 1.015625 P14 0.375 0.5625 0.53125 0.5 1.03125 P15 0.75 0.375 1.03125 0.75 2.5 P16 0.375 0.3125 0.5002 0.625 0.53125 P17 0.5 1.03125 1.0625 1.125 3 P18 0.5 0.625 0.375 1.0625 2.0625 P19 0.625 0.5078 0.5625 0.53125 0.625 P20 0.3125 0.5005 0.5625 0.625 0.625

TABLE P30 Distribution statistical results of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Range CAZ CFP AMK IMP LVX FIC ≤ 0.5 40% 20% 20% 15% 10% 0.5 < FIC ≤ 1 35% 55% 45% 55% 35% 1 < FIC ≤ 2 25% 25% 35% 30% 35% FIC > 2 — — — — 20%

TABLE P31 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 400 50 32 8 P2 200 100 64 8 P3 400 50 4 4 P4 400 50 256 64 P5 400 400 64 0.5 P6 400 100 512 256 P7 400 200 64 32 P8 800 50 256 64 P9 400 400 8 0.5 P10 400 400 4 0.5 P11 400 400 2 0.5 P12 400 200 64 0.5 P13 400 100 16 8 P14 800 100 16 4 P15 400 100 256 128 P16 800 100 32 8 P17 400 100 32 8 P18 400 100 64 16 P19 800 400 4 0.5 P20 800 50 256 64

TABLE P32 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the CFP and MIC after combined use EA EA CFP CFP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 400 50 128 32 P2 200 50 256 128 P3 400 400 16 0.5 P4 400 400 8 0.5 P5 400 50 512 256 P6 400 200 512 256 P7 400 50 512 256 P8 800 400 16 0.5 P9 400 50 64 32 P10 400 400 8 0.5 P11 400 400 8 0.5 P12 400 200 512 0.5 P13 400 100 256 32 P14 800 400 8 0.5 P15 400 50 64 16 P16 800 50 1024 256 P17 400 400 16 0.5 P18 400 50 256 128 P19 800 400 64 0.5 P20 800 400 1024 0.5

TABLE P33 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the AMK and MIC after combined use EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 400 400 1 0.0625 P2 200 100 1 0.0625 P3 400 100 4 1 P4 400 400 1 0.5 P5 400 400 2 0.0625 P6 400 400 256 32 P7 400 400 1 0.0625 P8 800 50 1 0.5 P9 400 200 1 0.0625 P10 400 50 4 1 P11 400 100 1 0.5 P12 400 50 32 8 P13 400 200 2 0.0625 P14 800 400 2 0.0625 P15 400 400 2 0.0625 P16 800 400 256 0.0625 P17 400 400 1 0.0625 P18 400 100 16 2 P19 800 400 1 0.0625 P20 800 50 2 1

TABLE P34 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 400 50 32 16 P2 200 100 64 2 P3 400 50 64 2 P4 400 400 32 2 P5 400 200 64 16 P6 400 400 32 2 P7 400 400 32 2 P8 800 400 64 2 P9 400 100 32 16 P10 400 200 64 16 P11 400 400 8 2 P12 400 200 32 2 P13 400 100 8 2 P14 800 200 32 8 P15 400 200 32 8 P16 800 400 16 2 P17 400 400 16 2 P18 400 400 32 2 P19 800 400 64 2 P20 800 100 4 2

TABLE P35 Sensibilization effect of the Pithecellobium clypearia Benth 60% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 400 50 32 4 P2 200 100 4 0.03125 P3 400 400 8 0.03125 P4 400 200 8 4 P5 400 400 1 0.03125 P6 400 400 0.5 0.03125 P7 400 400 32 0.03125 P8 800 400 1 0.03125 P9 400 200 4 2 P10 400 100 128 32 P11 400 25 2 4 P12 400 400 1 0.5 P13 400 400 2 0.03125 P14 800 800 32 1 P15 400 800 2 1 P16 800 25 32 16 P17 400 800 4 4 P18 400 25 4 8 P19 800 100 16 8 P20 800 100 4 2

TABLE P36 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + CAZ 8 64 EA + CFP 4 256 EA + AMK 0.0625 4 EA + IMP 2 16 EA + LVX 1 8

With the single use of the Pithecellobium clypearia Benth 60% ethanol extract to the MDRPA, the MIC₅₀ is 800 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 1600 μg/ml, and the MBC₉₀ is 1600 μg/ml.

For the 20 MDRPAs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the CAZ that the two drugs have no antagonistic effect, wherein 40% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 32 μg/ml to 8 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 256 μg/ml to 64 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the CFP to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CFP is reduced from single-use 64 μg/ml to 4 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 512 μg/ml to 256 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the AMK is reduced from single-use 1 μg/ml to 0.0625 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 32 μg/ml to 2 μg/ml and is reduced by 93.75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the IMP that the two drugs have no antagonistic effect, wherein 15% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 2 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

With the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the LVX to the 20 MDRPAs, there are four strains with FIC greater than 2 and has the antagonistic effect, wherein 10% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRPA, when the concentration of the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 8 μg/ml and is reduced by 75%.

2.4. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics (the LVX, the IMP, the AMK, the CAZ and the CFP) to the MDRPAs are shown in table P37.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRPAs are shown in table P38.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics to the MDRPAs is shown in table P39.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRPAs is shown in table P40.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics and distribution statistical results of the FIC values are shown in table P41 and table P42.

The sensibilization effects of the Pithecellobium clypearia Benth 95% ethanol extract to the five antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables P43-P48.

TABLE P37 In-vitro inhibitory test results of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics to the MDRPAs MIC(μg/ml) Strain No. EA LVX IMP AMK CAZ CFP P1 800 32 32 1 32 128 P2 800 4 64 1 64 256 P3 800 8 64 4 4 16 P4 800 8 32 1 256 8 P5 800 1 64 2 64 512 P6 800 0.5 32 256 512 512 P7 800 32 32 1 64 512 P8 800 1 64 1 256 16 P9 800 4 32 1 8 64 P10 800 128 64 4 4 8 P11 800 2 8 1 2 8 P12 800 1 32 32 64 512 P13 800 2 8 2 16 256 P14 800 32 32 2 16 8 P15 800 2 32 2 256 64 P16 800 32 16 256 32 >512 P17 800 4 16 1 32 16 P18 800 4 32 16 64 256 P19 800 16 64 1 4 64 P20 800 4 4 2 256 >512 ATCC27853 400 1 4 2 4 8

TABLE P38 In-vitro bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRPAs Strain No. MBC (μg/ml) P1 1600 P2 1600 P3 1600 P4 1600 P5 1600 P6 1600 P7 1600 P8 1600 P9 1600 P10 1600 P11 1600 P12 1600 P13 1600 P14 1600 P15 1600 P16 3200 P17 1600 P18 1600 P19 1600 P20 1600

TABLE P39 Statistical results on in-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics to the MDRPAs MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 800 800 800 CAZ 2-512 32 256 CFP 8-512 64 512 AMK 1-256 1 32 IMP 4-64  32 64 LVX 0.5-128  4 32

TABLE P40 Bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRPAs MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 1600-3200 1600 1600

TABLE P41 FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics Strain FIC No. EA + CAZ EA + CFP EA + AMK EA + IMP EA + LVX P1 1 0.5 0.5625 1 0.625 P2 0.75 1 0.3125 0.3125 0.507813 P3 0.75 0.5 1 0.3125 0.503906 P4 1 1 1.5 0.75 1 P5 1 1 1.03125 0.375 1.03125 P6 1 0.75 1.125 0.5625 1.0625 P7 0.75 0.515625 0.37 0.5625 1.000977 P8 0.5 0.28125 0.75 0.5 0.53125 P9 1 1 0.3125 1 1 P10 1 0.375 0.5 0.5 0.75 P11 1 1 1 0.75 1 P12 0.53125 0.503906 0.5 0.5625 1 P13 0.5 0.375 0.28125 0.75 1 P14 1 0.53125 0.53125 0.625 0.3125 P15 1 0.375 0.28125 0.375 1 P16 0.375 0.5 0.507813 0.625 0.625 P17 0.375 0.53125 0.5625 0.625 1 P18 1 0.75 0.5 0.375 1.5 P19 0.75 0.28125 0.5625 0.375 0.5 P20 0.5 0.500977 1 1 0.75

TABLE P42 Distribution statistical results of the FIC values of the combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the five antibiotics EA + EA + EA + EA + EA + FIC Strain No. CAZ CFP AMK IMP LVX FIC ≤ 0.5 25% 40% 40% 40% 10% 0.5 < FIC ≤ 1 75% 60% 45% 60% 70% 1 < FIC ≤ 2 — — 15% — 20% FIC > 2 — — — — —

TABLE P43 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the CAZ and MIC after combined use EA EA CAZ CAZ Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 16 P2 800 400 64 16 P3 800 400 4 1 P4 800 400 256 128 P5 800 400 64 32 P6 800 400 512 256 P7 800 200 64 32 P8 800 200 256 64 P9 800 400 8 4 P10 800 400 4 2 P11 800 400 2 1 P12 800 400 64 2 P13 800 200 16 4 P14 800 400 16 8 P15 800 400 256 128 P16 800 200 32 4 P17 800 200 32 4 P18 800 400 64 32 P19 800 400 4 1 P20 800 200 256 64

TABLE P44 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the CFP and MIC after combined use EA EA CFP CFP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 200 128 32 P2 800 400 256 128 P3 800 200 16 4 P4 800 400 8 4 P5 800 400 512 256 P6 800 400 512 128 P7 800 400 512 8 P8 800 200 16 0.5 P9 800 400 64 32 P10 800 200 8 1 P11 800 400 8 4 P12 800 400 512 2 P13 800 200 256 32 P14 800 400 8 0.25 P15 800 200 64 8 P16 800 200 1024 256 P17 800 400 16 0.5 P18 800 200 256 128 P19 800 200 64 2 P20 800 400 1024 1

TABLE P45 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the AMK and MIC after combined use EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 1 0.0625 P2 800 200 1 0.0625 P3 800 400 4 2 P4 800 800 1 0.5 P5 800 800 2 0.0625 P6 800 800 256 32 P7 800 200 1 0.12 P8 800 200 1 0.5 P9 800 200 1 0.0625 P10 800 200 4 1 P11 800 400 1 0.5 P12 800 200 32 8 P13 800 200 2 0.0625 P14 800 400 2 0.0625 P15 800 200 2 0.0625 P16 800 400 256 2 P17 800 400 1 0.0625 P18 800 200 16 4 P19 800 400 1 0.0625 P20 800 400 2 1

TABLE P46 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the IMP and MIC after combined use EA EA IMP IMP Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 16 P2 800 200 64 4 P3 800 200 64 4 P4 800 400 32 8 P5 800 200 64 8 P6 800 400 32 2 P7 800 400 32 2 P8 800 200 64 16 P9 800 400 32 16 P10 800 200 64 16 P11 800 200 8 4 P12 800 400 32 2 P13 800 400 8 2 P14 800 400 32 4 P15 800 200 32 4 P16 800 400 16 2 P17 800 400 16 2 P18 800 200 32 4 P19 800 200 64 8 P20 800 400 4 2

TABLE P47 Sensibilization effect of the Pithecellobium clypearia Benth 95% ethanol extract to the LVX and MIC after combined use EA EA LVX LVX Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) P1 800 400 32 4 P2 800 400 4 0.03125 P3 800 400 8 0.03125 P4 800 400 8 4 P5 800 800 1 0.03125 P6 800 800 0.5 0.03125 P7 800 800 32 0.03125 P8 800 400 1 0.03125 P9 800 400 4 2 P10 800 400 128 32 P11 800 400 2 1 P12 800 400 1 0.5 P13 800 400 2 1 P14 800 200 32 2 P15 800 400 2 1 P16 800 400 32 4 P17 800 400 4 2 P18 800 400 4 4 P19 800 200 16 4 P20 800 400 4 1

TABLE P48 MIC₅₀ and MIC₉₀ of the five antibiotics after combined use Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + CAZ 16 128 EA + CFP 4 128 EA + AMK 0.12 4 EA + IMP 4 16 EA + LVX 1 4

With the single use of the Pithecellobium clypearia Benth 95% ethanol extract to the MDRPA, the MIC₅₀ is 800 μg/ml, the MIC₉₀ is 800 μg/ml, the MBC₅₀ is 1600 μg/ml, and the MBC₉₀ is 1600 μg/ml.

For the 20 MDRPAs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the CAZ that the two drugs have no antagonistic effect, wherein 25% with FIC≤0.5 have the synergistic effect.

For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CAZ is reduced from single-use 32 μg/ml to 16 μg/ml and is reduced by 50%; and the MIC₉₀ is reduced from 256 μg/ml to 128 μg/ml and is reduced by 50%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the CFP to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 40% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the CFP is reduced from single-use 64 μg/ml to 4 μg/ml and is reduced by 93.75%; and the MIC₉₀ is reduced from 512 μg/ml to 128 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 40% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the AMK is reduced from single-use 1 μg/ml to 0.12 μg/ml and is reduced by 87.5%; and the MIC₉₀ is reduced from 32 μg/ml to 4 μg/ml and is reduced by 87.5%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the IMP that the two drugs have no antagonistic effect, wherein 40% with FIC≤0.5 have the synergistic effect. For the 20 MDRPAs, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the IMP is reduced from single-use 32 μg/ml to 4 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the LVX to the 20 MDRPAs that the two drugs have no antagonistic effect, wherein 10% with FIC≤0.5 show that the two drugs have a certain synergistic effect. For the 20 MDRPA, when the concentration of the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ of the LVX is reduced from single-use 4 μg/ml to 1 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 32 μg/ml to 4 μg/ml and is reduced by 87.5%.

III. Inhibitory and Bactericidal Tests of the ESBL-Producing Escherichia coli (Hereinafter Referred to as an ESBL-Producing ECO) Resistance of the EA and Test of the Sensibilization Effect by Respectively Using with the AMK or the SXT

Embodiment 3

1. Test Method: The MDRPA (Serial No.: E1-E20) Strains are Tested and are Evaluated with Reference to the Method in the First Embodiment.

2. Test Results

2.1. In-vitro inhibitory test results of the Pithecellobium clypearia Benth water extract and the two antibiotics (the AMK and the SXT) to the ESBL-producing ECOs are shown in table E1.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth water extract to the ESBL-producing ECOs are shown in table E2.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract and the two antibiotics to the ESBL-producing ECOs is shown in table E3.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the EA to the ESBL-producing ECOs is shown in table E4.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth water extract and the two antibiotics and distribution statistical results of the FIC values are shown in table E5 and table E6.

The sensibilization effects of the Pithecellobium clypearia Benth water extract to the two antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables E7-E9.

TABLE E1 MIC (μg/ml) Strain No. EA AMK SXT E1 1600 4 2432/128 E2 1600 16 2432/128 E3 1600 512 2432/128 E4 1600 4 76/4 E5 1600 16 2432/128 E6 1600 4 2432/128 E7 1600 32 2432/128 E8 1600 32 2432/128 E9 1600 64 2432/128 E10 1600 8 2432/128 E11 1600 32 76/4 E12 1600 32 2432/128 E13 1600 4 2432/128 E14 1600 8 2432/128 E15 1600 8 2432/128 E16 1600 16 2432/128 E17 1600 8 2432/128 E18 1600 128 2432/128 E19 1600 64 152/8  E20 1600 16 76/4 ATCC25922 800 4  9.5/0.5

TABLE E2 Strain No. MBC(μg/ml) E1 3200 E2 3200 E3 3200 E4 3200 E5 3200 E6 3200 E7 3200 E8 3200 E9 3200 E10 3200 E11 3200 E12 3200 E13 3200 E14 3200 E15 3200 E16 3200 E17 3200 E18 3200 E19 3200 E20 3200

TABLE E3 MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 1600 1600 1600 AMK 4-512  16 64 SXT 76/4-2432/128 2432/128 2432/128

TABLE E4 MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 3200 3200 3200

TABLE E5 FIC Strain No. EA + AMK EA + SXT E1 0.75 0.507813 E2 0.375 1.003906 E3 0.28125 0.507813 E4 0.75 0.75 E5 0.625 0.507813 E6 1 0.515625 E7 0.375 0.507813 E8 0.5 0.507813 E9 0.375 0.500061 E10 0.75 0.507813 E11 0.5 0.375 E12 0.75 0.125485 E13 1.25 0.507813 E14 0.75 0.507813 E15 0.75 0.500058 E16 0.5 0.125485 E17 0.5 0.507813 E18 0.625 0.250977 E19 0.375 0.375 E20 0.5 0.75

TABLE E6 FIC Range EA + AMK EA + SXT FIC ≤ 0.5 50% 25% 0.5 < FIC ≤ 1 45% 70% 1 < FIC ≤ 2  5%  5% FIC > 2 — —

TABLE E7 EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 400 4 2 E2 1600 200 16 4 E3 1600 400 512 16 E4 1600 400 4 2 E5 1600 200 16 8 E6 1600 800 4 2 E7 1600 200 32 8 E8 1600 400 32 8 E9 1600 400 64 8 E10 1600 400 8 4 E11 1600 400 32 8 E12 1600 400 32 16 E13 1600 800 4 2 E14 1600 400 8 4 E15 1600 400 8 4 E16 1600 400 16 4 E17 1600 400 8 2 E18 1600 800 128 16 E19 1600 200 64 16 E20 1600 400 16 4

TABLE E8 EA EA SXT SXT Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 800 2432/128 19/1 E2 1600 1600 2432/128  9.5/0.5 E3 1600 800 2432/128 19/1 E4 1600 800 76/4 19/1 E5 1600 800 2432/128 19/1 E6 1600 800 2432/128 38/2 E7 1600 800 2432/128 19/1 E8 1600 800 2432/128 19/1 E9 1600 800 2432/128  0.148438/0.078125 E10 1600 800 2432/128 19/1 E11 1600 200 76/4 19/1 E12 1600 200 2432/128  1.1875/0.0625 E13 1600 800 2432/128 19/1 E14 1600 800 2432/128 19/1 E15 1600 800 2432/128  0.148438/0.078125 E16 1600 200 2432/128  1.1875/0.0625 E17 1600 800 2432/128 19/1 E18 1600 400 2432/128  2.375/0.125 E19 1600 400 152/8  19/1 E20 1600 800 76/4 19/1

TABLE E9 Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + AMK 4 16 EA + SXT 19/1 19/1

The test results shows that, with the single use of the Pithecellobium clypearia Benth water extract to the ESBL-producing ECOs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 3200 μg/ml, and the MBC₉₀ is 3200 μg/ml.

For the 20 ESBL-producing ECOs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth water extract and the AMK that the two drugs have no antagonistic effect, wherein 50% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth water extract is smaller than or equal to ½ MIC, the MIC₅₀ of the AMK is reduced from single-use 16 μg/ml to 4 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth water extract and the SXT to the 20 ESBL-producing ECOs that the two drugs have no antagonistic effect, wherein 25% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth water extract is smaller than or equal to the single-use MIC, the MIC₅₀ and the MIC₉₀ of the SXT are respectively reduced from single-use 2432/128 μg/ml to 19/1 μg/ml, and are reduced by 99.3%.

2.2. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 10% ethanol extract and the two antibiotics (the AMK and the SXT) to the ESBL-producing ECOs are shown in table E10.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing ECOs are shown in table E11.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract and the two antibiotics to the ESBL-producing ECOs is shown in table E12.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing ECOs is shown in table E13.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 10% ethanol extract and the two antibiotics and distribution statistical results of the FIC values are shown in table E14 and table E15.

The sensibilization effects of the Pithecellobium clypearia Benth 10% ethanol extract to the two antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables E16-E18.

TABLE E10 MIC (μg/ml) Strain No. EA AMK SXT E1 1600 4 2432/128 E2 1600 16 2432/128 E3 1600 512 2432/128 E4 1600 4 76/4 E5 1600 16 2432/128 E6 1600 4 2432/128 E7 1600 32 2432/128 E8 1600 32 2432/128 E9 1600 64 2432/128 E10 1600 8 2432/128 E11 1600 32 76/4 E12 1600 32 2432/128 E13 1600 4 2432/128 E14 1600 8 2432/128 E15 1600 8 2432/128 E16 1600 16 2432/128 E17 1600 8 2432/128 E18 1600 128 2432/128 E19 1600 64 152/8  E20 1600 16 76/4 ATCC25922 800 4  9.5/0.5

TABLE E11 Strain No. MBC(μg/ml) E1 3200 E2 3200 E3 3200 E4 3200 E5 3200 E6 3200 E7 3200 E8 3200 E9 3200 E10 3200 E11 3200 E12 3200 E13 3200 E14 3200 E15 3200 E16 3200 E17 3200 E18 3200 E19 3200 E20 3200

TABLE E12 MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 1600 1600 1600 AMK 4-512  16 64 SXT 76/4-2432/128 2432/128 2432/128

TABLE E13 MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 3200 3200 3200

TABLE E14 FIC Strain No. EA + AMK EA + SXT E1 1 0.507813 E2 0.75 0.507813 E3 0.265625 0.507813 E4 0.5 0.75 E5 0.5 0.507813 E6 1 0.507813 E7 0.5 0.507813 E8 0.75 0.507813 E9 0.5 0.500061 E10 0.75 0.507813 E11 0.5 0.5 E12 0.75 0.250975 E13 1 0.507813 E14 1 0.507813 E15 0.75 0.500058 E16 0.5 0.125485 E17 0.75 0.507813 E18 0.5 1.000977 E19 0.375 0.375 E20 0.5 0.75

TABLE E15 FIC Range EA + AMK EA + SXT FIC ≤ 0.5 50% 20% 0.5 < FIC ≤ 1 50% 75% 1 < FIC ≤ 2 —  5% FIC > 2 — —

TABLE E16 EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 800 4 2 E2 1600 800 16 4 E3 1600 400 512 8 E4 1600 400 4 1 E5 1600 400 16 4 E6 1600 800 4 2 E7 1600 400 32 8 E8 1600 800 32 8 E9 1600 400 64 16 E10 1600 400 8 4 E11 1600 400 32 8 E12 1600 400 32 16 E13 1600 800 4 2 E14 1600 800 8 4 E15 1600 400 8 4 E16 1600 400 16 4 E17 1600 800 8 2 E18 1600 400 128 32 E19 1600 400 64 8 E20 1600 400 16 4

TABLE E17 EA EA SXT SXT Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 800 2432/128 19/1 E2 1600 800 2432/128 19/1 E3 1600 800 2432/128 19/1 E4 1600 800 76/4 19/1 E5 1600 800 2432/128 19/1 E6 1600 800 2432/128 19/1 E7 1600 800 2432/128 19/1 E8 1600 800 2432/128 19/1 E9 1600 800 2432/128  0.148438/0.078125 E10 1600 800 2432/128 19/1 E11 1600 400 76/4 19/1 E12 1600 400 2432/128  2.375/0.125 E13 1600 800 2432/128 19/1 E14 1600 800 2432/128 19/1 E15 1600 800 2432/128  0.148438/0.078125 E16 1600 200 2432/128  1.1875/0.0625 E17 1600 800 2432/128 19/1 E18 1600 1600 2432/128  2.375/0.125 E19 1600 400 152/8  19/1 E20 1600 800 76/4 19/1

TABLE E18 Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + AMK 4 16 EA + SXT 19/1 19/1

The test results shows that, with the single use of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing ECOs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 3200 μg/ml, and the MBC₉₀ is 3200 μg/ml.

For the 20 ESBL-producing ECOs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 50% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to ½ MIC, the MIC₅₀ of the AMK is reduced from single-use 16 μg/ml to 4 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 10% ethanol extract and the SXT to the 20 ESBL-producing ECOs that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 10% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ and the MIC₉₀ of the SXT are respectively reduced from single-use 2432/128 μg/ml to 19/1 μg/ml, and are reduced by 99.3%.

2.3. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 60% ethanol extract and the two antibiotics (the AMK and the SXT) to the ESBL-producing ECOs are shown in table E19.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing ECOs are shown in table E20.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract and the two antibiotics to the ESBL-producing ECOs is shown in table E21.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing ECOs is shown in table E22.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 60% ethanol extract and the two antibiotics and distribution statistical results of the FIC values are shown in table E23 and table E24.

The sensibilization effects of the Pithecellobium clypearia Benth 60% ethanol extract to the two antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables E25-E27.

TABLE E19 MIC (μg/ml) Strain No. EA AMK SXT E1 800 4 2432/128 E2 800 16 2432/128 E3 800 512 2432/128 E4 800 4 76/4 E5 800 16 2432/128 E6 1600 4 2432/128 E7 800 32 2432/128 E8 800 32 2432/128 E9 800 64 2432/128 E10 800 8 2432/128 E11 800 32 76/4 E12 800 32 2432/128 E13 400 4 2432/128 E14 800 8 2432/128 E15 800 8 2432/128 E16 800 16 2432/128 E17 800 8 2432/128 E18 800 128 2432/128 E19 800 64 152/8  E20 800 16 76/4 ATCC25922 800 4  9.5/0.5

TABLE E20 Strain No. MBC(μg/ml) E1 1600 E2 1600 E3 1600 E4 1600 E5 1600 E6 >1600 E7 1600 E8 1600 E9 1600 E10 1600 E11 1600 E12 1600 E13 >1600 E14 1600 E15 1600 E16 1600 E17 1600 E18 1600 E19 1600 E20 1600

TABLE E21 MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 400-1600   800 800 AMK 4-512  16 64 SXT 76/4-2432/128 2432/128 2432/128

TABLE E22 MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA ≥1600 1600 1600

TABLE E23 FIC Strain no. EA + AMK EA + SXT E1 0.5625 1.0078 E2 0.25 1.0039 E3 0.2656 1.0001 E4 0.5625 0.75 E5 0.5625 0.5078 E6 0.375 0.2656 E7 0.125 0.5078 E8 0.1875 0.5078 E9 0.3125 1.0001 E10 0.5625 0.5078 E11 0.1875 0.0703 E12 0.1875 0.0630 E13 1.125 1.0039 E14 0.5625 0.5156 E15 0.3125 1.0001 E16 0.1875 0.0630 E17 0.3125 0.5078 E18 0.375 0.0635 E19 0.25 0.625 E20 0.3125 0.375

TABLE E24 FIC Range EA + AMK EA + SXT FIC ≤ 0.5 70% 30% 0.5 < FIC ≤ 1 25% 40% 1 < FIC ≤ 2  5% 30% FIC > 2 — —

TABLE E25 EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 800 50 4 2 E2 800 100 16 2 E3 800 200 512 8 E4 800 50 4 2 E5 800 50 16 8 E6 1600 400 4 0.5 E7 800 50 32 2 E8 800 50 32 4 E9 800 200 64 4 E10 800 50 8 4 E11 800 50 32 4 E12 800 50 32 4 E13 400 50 4 4 E14 800 50 8 4 E15 800 50 8 2 E16 800 50 16 2 E17 800 50 8 2 E18 800 100 128 32 E19 800 100 64 8 E20 800 50 16 4

TABLE E26 EA EA SXT SXT Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 800 800 2432/128 19/1 E2 800 800 2432/128  9.5/0.5 E3 800 800 2432/128  0.148438/0.078125 E4 800 400 76/4 19/1 E5 800 400 2432/128 19/1 E6 1600 400 2432/128 38/2 E7 800 400 2432/128 19/1 E8 800 400 2432/128 19/1 E9 800 800 2432/128  0.148438/0.078125 E10 800 400 2432/128 19/1 E11 800 50 76/4 19/1 E12 800 50 2432/128  1.1875/0.0625 E13 400 400 2432/128  9.5/0.5 E14 800 400 2432/128 38/2 E15 800 800 2432/128  0.148438/0.078125 E16 800 50 2432/128  1.1875/0.0625 E17 800 400 2432/128 19/1 E18 800 50 2432/128  2.375/0.125 E19 800 400 152/8  19/1 E20 800 100 76/4 19/1

TABLE E27 Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + AMK 4 8 EA + SXT 19/1 19/1

The test results shows that, with the single use of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing ECOs, the MIC₅₀ is 800 μg/ml, the MIC₉₀ is 800 μg/ml, the MBC₅₀ is 1600 μg/ml, and the MBC₉₀ is 1600 μg/ml.

For the 20 ESBL-producing ECOs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 70% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to ¼ MIC, the MIC₅₀ of the AMK is reduced from single-use 16 μg/ml to 4 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 64 μg/ml to 8 μg/ml.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 60% ethanol extract and the SXT to the 20 ESBL-producing ECOs that the two drugs have no antagonistic effect, wherein 30% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 60% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ and the MIC₉₀ of the SXT are respectively reduced from single-use 2432/128 μg/ml to 19/1 μg/ml, and are reduced by 99.3%.

2.4. In-vitro inhibitory test results of the Pithecellobium clypearia Benth 95% ethanol extract and the two antibiotics (the AMK and the SXT) to the ESBL-producing ECOs are shown in table E28.

In-vitro bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing ECOs are shown in table E29.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract and the two antibiotics to the ESBL-producing ECOs is shown in table E30.

The statistic analysis on bactericidal MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing ECOs is shown in table E31.

FIC values of a combined drug sensitive test of the Pithecellobium clypearia Benth 95% ethanol extract and the two antibiotics and distribution statistical results of the FIC values are shown in table E32 and table E33.

The sensibilization effects of the Pithecellobium clypearia Benth 95% ethanol extract to the two antibiotics and the MIC₅₀ and MIC₉₀ after combined use are shown in tables E34-E36.

TABLE E28 MIC (μg/ml) Strain No. EA AMK SXT E1 1600 4 2432/128 E2 1600 16 2432/128 E3 1600 512 2432/128 E4 1600 4 76/4 E5 1600 16 2432/128 E6 1600 4 2432/128 E7 1600 32 2432/128 E8 1600 32 2432/128 E9 1600 64 2432/128 E10 1600 8 2432/128 E11 1600 32 76/4 E12 1600 32 2432/128 E13 1600 4 2432/128 E14 1600 8 2432/128 E15 1600 8 2432/128 E16 1600 16 2432/128 E17 1600 8 2432/128 E18 1600 128 2432/128 E19 1600 64 152/8  E20 1600 16 76/4 ATCC25922 800 4  9.5/0.5

TABLE E29 Strain No. MBC(μg/ml) E1 3200 E2 3200 E3 3200 E4 3200 E5 3200 E6 3200 E7 3200 E8 3200 E9 3200 E10 3200 E11 3200 E12 3200 E13 3200 E14 3200 E15 3200 E16 3200 E17 3200 E18 3200 E19 3200 E20 3200

TABLE E30 MIC (μg/ml) Drug Range MIC₅₀ MIC₉₀ EA 1600 1600 1600 AMK 4-512  16 64 SXT 76/4-2432/128 2432/128 2432/128

TABLE E31 MBC (μg/ml) Drug Range MBC₅₀ MBC₉₀ EA 3200 3200 3200

TABLE E32 FIC Strain No. EA + AMK EA + SXT E1 1 0.507813 E2 0.5 0.507813 E3 0.53125 0.507813 E4 0.5 0.75 E5 0.5 0.507813 E6 0.75 0.507813 E7 0.75 0.503906 E8 0.75 0.507813 E9 0.75 0.503906 E10 0.75 0.507813 E11 0.75 0.75 E12 0.5 1.000975 E13 0.75 0.507813 E14 0.75 0.507813 E15 0.75 0.500058 E16 0.5 0.132813 E17 0.75 0.507813 E18 0.5 1.000977 E19 0.375 0.625 E20 0.5 0.75

TABLE E33 FIC Range EA + AMK EA + SXT FIC ≤ 0.5 40%  5% 0.5 < FIC ≤ 1 60% 85% 1 < FIC ≤ 2 — 10% FIC > 2 — —

TABLE E34 EA EA AMK AMK Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 800 4 2 E2 1600 400 16 4 E3 1600 800 512 16 E4 1600 400 4 1 E5 1600 400 16 4 E6 1600 800 4 1 E7 1600 800 32 8 E8 1600 800 32 8 E9 1600 800 64 16 E10 1600 400 8 4 E11 1600 800 32 8 E12 1600 400 32 8 E13 1600 400 4 2 E14 1600 800 8 2 E15 1600 400 8 4 E16 1600 400 16 4 E17 1600 800 8 2 E18 1600 400 128 32 E19 1600 400 64 8 E20 1600 400 16 4

TABLE E35 EA EA SXT SXT Strain single-use combined-use single-use combined-use No. MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) MIC (μg/ml) E1 1600 800 2432/128 19/1 E2 1600 800 2432/128 19/1 E3 1600 800 2432/128 19/1 E4 1600 800 76/4 19/1 E5 1600 800 2432/128 19/1 E6 1600 800 2432/128 19/1 E7 1600 800 2432/128  9.5/0.5 E8 1600 800 2432/128 19/1 E9 1600 800 2432/128  9.5/0.5 E10 1600 800 2432/128 19/1 E11 1600 800 76/4 19/1 E12 1600 1600 2432/128  2.375/0.125 E13 1600 800 2432/128 19/1 E14 1600 800 2432/128 19/1 E15 1600 800 2432/128  0.148438/0.078125 E16 1600 200 2432/128 19/1 E17 1600 800 2432/128 19/1 E18 1600 1600 2432/128  2.375/0.125 E19 1600 800 152/8  19/1 E20 1600 800 76/4 19/1

TABLE E36 Antibiotic (μg/ml) Combined drug MIC₅₀ after combined use MIC₉₀ after combined use EA + AMK 4 16 EA + SXT 19/1 19/1

The test results shows that, with the single use of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing ECOs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, the MBC₅₀ is 3200 μg/ml, and the MBC₉₀ is 3200 μg/ml.

For the 20 ESBL-producing ECOs, it is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the AMK that the two drugs have no antagonistic effect, wherein 50% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to ½ MIC, the MIC₅₀ of the AMK is reduced from single-use 16 μg/ml to 4 μg/ml and is reduced by 75%; and the MIC₉₀ is reduced from 64 μg/ml to 16 μg/ml and is reduced by 75%.

It is indicated by FIC≤2 with the combined use of the Pithecellobium clypearia Benth 95% ethanol extract and the SXT to the 20 ESBL-producing ECOs that the two drugs have no antagonistic effect, wherein 20% with FIC≤0.5 have the synergistic effect. When the Pithecellobium clypearia Benth 95% ethanol extract is smaller than or equal to the single-use MIC, the MIC₅₀ and the MIC₉₀ of the SXT are respectively reduced from single-use 2432/128 μg/ml to 19/1 μg/ml, and are reduced by 99.3%.

IV. Inhibitory and Bactericidal Tests of ESBL-Producing Klebsiella pneumoniae (Hereinafter Referred to as an ESBL-Producing KPN) Resistance of the EA

Embodiment 4

1. Test Method: The ESBL-Producing KPN (Serial No.: K1-K20) Strains are Tested and are Evaluated with Reference to the Inhibitory and Bactericidal Test Method in the First Embodiment.

2. Test Results

In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs are shown in table K1.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀, MIC₉₀, MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs is shown in table K2 and table K3.

In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing KPNs are shown in table K4.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀, MIC₉₀, MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing KPNs is shown in table K5 and table K6.

In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs are shown in table K7.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀, MIC₉₀, MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs is shown in table K8 and table K9.

In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs are shown in table K10.

The statistic analysis on in-vitro inhibitory and bactericidal MIC₅₀, MIC₉₀, MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs is shown in table K11 and table K12.

TABLE K1 In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs Strain No. MIC(μg/ml) MBC(μg/ml) K1 1600 >1600 K2 1600 >1600 K3 1600 >1600 K4 1600 >1600 K5 1600 >1600 K6 1600 >1600 K7 1600 >1600 K8 1600 >1600 K9 1600 >1600 K10 1600 >1600 K11 1600 >1600 K12 1600 >1600 K13 1600 >1600 K14 1600 >1600 K15 1600 >1600 K16 1600 >1600 K17 1600 >1600 K18 1600 >1600 K19 1600 >1600 K20 1600 >1600

TABLE K2 In-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs MIC Range MIC₅₀ MIC₉₀ 1600 1600 1600

TABLE K3 In-vitro inhibitory MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs MBC Range MBC₅₀ MBC₉₀ >1600 >1600 >1600

TABLE K4 In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing KPNs Strain No. MIC(μg/ml) MBC(μg/ml) K1 1600 >1600 K2 800 >1600 K3 1600 >1600 K4 800 >1600 K5 1600 >1600 K6 1600 >1600 K7 1600 >1600 K8 1600 >1600 K9 1600 >1600 K10 1600 >1600 K11 1600 >1600 K12 1600 >1600 K13 1600 >1600 K14 1600 >1600 K15 800 >1600 K16 800 >1600 K17 1600 >1600 K18 1600 >1600 K19 1600 >1600 K20 1600 >1600

TABLE K5 In-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing KPNs MIC Range MIC₅₀ MIC₉₀ 800-1600 1600 1600

TABLE K6 In-vitro inhibitory MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 10% ethanol extract to the ESBL-producing KPNs MBC Range MBC₅₀ MBC₉₀ >1600 >1600 >1600

TABLE K7 In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs Strain No. MIC(μg/ml) MBC(μg/ml) K1 1600 >1600 K2 800 >1600 K3 800 >1600 K4 800 >1600 K5 1600 >1600 K6 1600 >1600 K7 400 >1600 K8 800 >1600 K9 1600 >1600 K10 1600 >1600 K11 1600 >1600 K12 1600 >1600 K13 800 >1600 K14 800 >1600 K15 400 >1600 K16 800 >1600 K17 1600 >1600 K18 1600 >1600 K19 800 >1600 K20 800 >1600

TABLE K8 In-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs MIC Range MIC₅₀ MIC₉₀ 400-1600 800 1600

TABLE K9 In-vitro inhibitory MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs MBC Range MBC₅₀ MBC₉₀ >1600 >1600 >1600

TABLE K10 In-vitro inhibitory and bactericidal test results of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs Strain No. MIC(μg/ml) MBC(μg/ml) K1 1600 >1600 K2 1600 >1600 K3 1600 >1600 K4 1600 >1600 K5 1600 >1600 K6 1600 >1600 K7 1600 >1600 K8 1600 >1600 K9 1600 >1600 K10 1600 >1600 K11 1600 >1600 K12 1600 >1600 K13 800 >1600 K14 800 >1600 K15 1600 >1600 K16 1600 >1600 K17 1600 >1600 K18 1600 >1600 K19 1600 >1600 K20 1600 >1600

TABLE K11 In-vitro inhibitory MIC₅₀ and MIC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs MIC Range MIC₅₀ MIC₉₀ 800-1600 1600 1600

TABLE K12 In-vitro inhibitory MBC₅₀ and MBC₉₀ of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs MBC Range MBC₅₀ MBC₉₀ >1600 >1600 >1600

With the single use of the Pithecellobium clypearia Benth water extract to the ESBL-producing KPNs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, and both the MBC₅₀ and the MBC₉₀ are greater than 1600 μg/ml.

With the single use of the Pithecellobium clypearia Benth 10% ethanol extract to the KPNs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, and both the MBC₅₀ and the MBC₉₀ are greater than 1600 μg/ml.

With the single use of the Pithecellobium clypearia Benth 60% ethanol extract to the ESBL-producing KPNs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, and both the MBC₅₀ and the MBC₉₀ are greater than 1600 μg/ml.

With the single use of the Pithecellobium clypearia Benth 95% ethanol extract to the ESBL-producing KPNs, the MIC₅₀ is 1600 μg/ml, the MIC₉₀ is 1600 μg/ml, and both the MBC₅₀ and the MBC₉₀ are greater than 1600 μg/ml. 

What is claimed is:
 1. A method of treating a disease caused by an Extended-Spectrum Beta-Lactamase (ESBL) producing Klebsiella pneumonia (KPN) resistant infection, comprising administering an ESBL-producing KPN resistant drug comprising a Pithecellobium clypearia Benth Extract (EA).
 2. The method of claim 1, wherein the EA is a Pithecellobium clypearia Benth water extract or a Pithecellobium clypearia Benth ethanol extract.
 3. The method of claim 2, wherein the Pithecellobium clypearia Benth water extract or the Pithecellobium clypearia Benth ethanol extract is prepared with a following method: extracting Pithecellobium clypearia Benth coarse powder with water or an ethanol aqueous solution having a concentration of 10%-95% by a volume ratio, and then extracting an obtained extracting solution with ethyl acetate, in which the obtained extract is a final product.
 4. The method of claim 3, wherein the ethanol aqueous solution is an ethanol aqueous solution having the concentration of 60% by the volume ratio. 